Microprobe, Inc.

Nicolaas M Vermuelen

Jeffrey Van Ness

Dennis E Schwartz

B Melina Cimler

Roy H Kanemoto

Trevor H Adams

This invention provides for a method of quantifying bacteria using a bacterial specific nucleic acid probe which is complementary to a unique and highly conserved region of the 16S ribosomal RNA (rRNA) of bacteria. This probe permits the rapid detection of 16S rRNA in a sample and by comparison with known standards, one can estimate the total bacterial count in the sample. The method is accurate and reproducible and conducted at temperatures of between about 120.degree. to about 40.degree. C.

Quantification of bacteria using a nucleic acid hybridization assay

This invention relates to novel methods for the extraction of nucleic acid. In particular methods are described for isolating nucleic acid from a sample containing a complex biological mixture of nucleic acid and non-nucleic acids wherein the sample is combined with an extraction solution comprising a lactam and then the nucleic acid material is isolated from the resulting combined solution. The resulting combined solution is mixed and becomes biphasic and the nucleic acid material is isolated from the aqueous phase by precipitation with ethanol. The lactam is preferably about 5 to about 70% of the extraction solution and is most preferably 2-pyrrolidone, N-ethyl-2-pyrrolidone, N-cyclohexyl-2-pyrrolidone, N-dodecyl-2-pyrrolidone, N-methyl-2-pyrrolidone, N-hydroxyethyl-2-pyrrolidone, N-methyl-2-piperidone, 2-.epsilon.-caprolactam, N-methyl-2-caprolactam, 2-piperidone or N-(4-hydroxybenzyl)pyrrolidone. Methods for selectively isolating DNA, ribosomal RNA and plasmid DNA are also disclosed.

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Woodinville, WA, United States of America

Nicolaas M Vermuelen